Figure 2. SDS PAGE induction expression and gel shift assay for DNA-binding activities in vitro.

a: Induction expression and purification of target proteins, GmZF1::GST fusion protein and GST protein. Lane 1-uninduced strains containing pGEX4T-1 vector; Lane 2-induced strains containing pGEX4T-1; Lane-3-uninduced strains containing GmZF1::GST fusion; Lane 4-induced strains containing GmZF1::GST fusion; Lane 5-purified target protein as indicated by arrows. b: Probe design and composition. Nucleotide sequences of EP1S and mutated EP1S (E1, E2 and E3) probes. The nucleotide mutations in the EP1S core motif for each probe are boxed. c: The gel shift assay was performed in a solution containing 0.2 µg GmZF1::GST proteins or GST proteins and the probes ³²P-labeled EP1S (E1) or mutant EP1S (E2 and E3), respectively. The GmZF1::EP1S complex and free probes are indicated by arrows.